Redistribution of fibronectin and cytoskeletal proteins during the differentiation of rat mammary tumor cells in vitro

Warburton, Michael J.; Head, Linda P.; Rudland, Philip S.

doi:10.1016/0014-4827(81)90082-3

Cited by 39 publications

(14 citation statements)

References 18 publications

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“…However, when time-lapse nine out of the ten major polypeptides that change in going from cuboidal epithelial to elongated, myoepithelial-like cells also change in approximately the same proportions in ring clones of elongated cells obtained from colchicine-treated Rama 25 cuboidal cells. The colchicine-generated elongated cell clones also show a large increase in deposition of the basement membrane proteins laminin, type IV collagen, and fibronectin and the rearrangement of the actomyosin cytoskeleton when observed by immunofluorescent techniques (unpublished results), similar to the changes seen in Rama 29 cells and in the uncloned "floaters" (Warburton et al, 1981(Warburton et al, , 1982. These results above, coupled with the fact that no new polypeptides are seen in the colchicine-generated elongated cell clones, would suggest that the colchicinegenerated elongated cells are virtually identical to the elongated, myoepithelial-like cells that occur spontaneously in Rama 25 cuboidal epithelial cultures.…”

Section: -Rsupporting

confidence: 56%

Microtubule‐disrupting drugs increase the frequency of conversion of a rat mammary epithelial stem cell line to elongated, myoepithelial‐like cells in culture

Paterson

Rudland

1985

Journal Cellular Physiology

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Rat mammary (Rama) 25 cuboidal epithelial stem cells convert at a low frequency to elongated, Thy-1-positive, myoepithelial-like cells in culture; one such cell line is termed Rama 29. Addition of increasing concentrations of the microtubule-disrupting drug colchicine to sparse cultures of Rama 25 dramatically increases the percentage of colonies containing elongated cells and the percentage of Thy-1-positive cells when the drug is removed. Similar results on the formation of elongated cell colonies are obtained with other microtubule disruptors, such as vinblastine, vincristine, demecolcine, and nocodazole. The inactive analogues of colchicine beta- and delta-lumicolchicine and the microfilamental-disruptors cytochalasin B and D are without effect on the formation of elongated cell colonies and Thy-1-positive cells. For a given concentration of colchicine the percentage of elongated cell colonies and Thy-1-positive cells increases the longer the cells are exposed to the drug (range 8-96 hr) and the longer the drug-treated cultures are subsequently grown in drug-free medium. Colchicine fails to display this morphological change on Rama 29 elongated cells and on Rama 600 epithelial cells from a rat mammary metastasizing tumor. Immunofluorescent localization of antisera to tubulin confirms that colchicine disrupts the microtubules in all three cell lines at similar concentrations (0.1 to 1 microM) to those required to increase the percentage of elongated cell colonies in Rama 25. The DNA synthesis inhibitor cytosine arabinoside fails to inhibit this conversion process, and time-lapse cinematographic studies confirm that the conversion of a cuboidal to an elongated cell can take place without cell division. However, cell division may sometimes be required for subsequent stabilization events. Treatment of Rama 25 cells with colchicine under the same conditions also increases the abundance of elongated cell (Rama 29)-associated polypeptides, and elongated cell clones isolated after such treatment show an overall pattern of protein synthesis very similar to that of Rama 29.

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Section: -Rsupporting

confidence: 56%

Microtubule‐disrupting drugs increase the frequency of conversion of a rat mammary epithelial stem cell line to elongated, myoepithelial‐like cells in culture

Paterson

Rudland

1985

Journal Cellular Physiology

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“…Formations of spontaneous dome structures in BuMECs grown on plastic substratum suggest that these cells undergo contact mediated differentiation and secrete basement membrane components. MEC lines have been reported to show dome-like structures in bovine [22], ovine (NISH) [14], human [33] and in rat in the presence of DMSO [34]. The SV40 larger T antigen induced immortalized MAC-T cells of bovine origin have also been reported to form dome-like structures when cultured on collagen [18] or when co-cultured with bovine myoepithelial cells [35].…”

Section: Discussionmentioning

confidence: 99%

Establishment and Characterization of a Buffalo (Bubalus bubalis) Mammary Epithelial Cell Line

Anand

Dogra²,

Singh

et al. 2012

PLoS ONE

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BackgroundThe objective of this study was to establish the buffalo mammary epithelial cell line (BuMEC) and characterize its mammary specific functions.MethodologyBuffalo mammary tissue collected from the slaughter house was processed enzymatically to obtain a heterogenous population of cells containing both epithelial and fibroblasts cells. Epithelial cells were purified by selective trypsinization and were grown in a plastic substratum. The purified mammary epithelial cells (MECs) after several passages were characterized for mammary specific functions by immunocytochemistry, RT-PCR and western blot.Principal FindingsThe established buffalo mammary epithelial cell line (BuMEC) exhibited epithelial cell characteristics by immunostaining positively with cytokeratin 18 and negatively with vimentin. The BuMEC maintained the characteristics of its functional differentiation by expression of β-casein, κ-casein, butyrophilin and lactoferrin. BuMEC had normal growth properties and maintained diploid chromosome number (2n = 50) before and after cryopreservation. A spontaneously immortalized buffalo mammary epithelial cell line was established after 20 passages and was continuously subcultured for more than 60 passages without senescence.ConclusionsWe have established a buffalo mammary epithelial cell line that can be used as a model system for studying mammary gland functions.

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“…Similar morphological forms have also been generated from mouse mammary tumor epithelial cells [93,94]. The elongated rat cells have been adjudged to be related to myoepithelial cells based on their ultrastructure and their immunocytochemical staining pattern [95][96][97]. However, like the elongated, myoepithelial-like cells in primary cultures of rat mammary glands [84], although the elongated cells always express vimentin, laminin, type IV collagen, fibronectin, Thy-1 [22, 91, 96--98] and the receptors for GS-1 and PWM lectins [19], the staining pattern with antisera to actin, myosin, and human keratin is variable; some clones stain well and others poorly, e.g., Rama 29 [99].…”

Section: (5) Culture Of Normal Rat Mammary Glands and Benign Tumorsmentioning

confidence: 94%

Stem cells and the development of mammary cancers in experimental rats and in humans

Rudland

1987

Cancer Metast Rev

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Evidence based on immunocytochemical staining and ultrastructure suggests that morphological gradations between epithelial and myoepithelial cells, and possibly between epithelial cells and alveolar-like cells occur in terminal ductal structures of rat and human mammary glands. The benign carcinogen-induced rat and benign human mammary tumors can contain epithelial, myoepithelial-like and alveolar-like cells, whereas the malignant counterparts mainly contain only epithelial-like cells. Clonal epithelial cell lines from normal rat mammary glands, benign tumors, and SV40-transformed human mammary glands can differentiate to either myoepithelial-like or alveolar-like cells. In those of the rat, the differentiation processes occur in steps: intermediate cells along the myoepithelial-like pathway resemble intermediates in terminal ductal structures in vivo, and can also generate certain well-differentiated mesenchymal elements of the original tumours. Differentiation of the benign rat cells to alveolar-like cells with mammatrophic hormones and retinoids in vitro leads to a reduction in their tumor-forming ability in vivo. Cell lines from malignant rat mammary tumors of increasing metastatic potential and from human ductal carcinomas largely fail to yield myoepithelial-like or alveolar-like cells and are relatively slow-growing. Growth of the rat mammary epithelial cells in culture is stimulated by a pituitary-derived mammatrophic growth factor (PMGF), prostaglandin E2, and alpha-transforming growth factor; the response of the malignant cell lines to PMGF is reduced. It is suggested that stem cells exist in the rat and human glands that are capable of differentiating to the other major cell types of the mammary parenchyma, and that during the carcinogenic process they generate genetically unstable cells which lose their ability to differentiate and attempt to maximise their intrinsically slow growth rate.

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Redistribution of fibronectin and cytoskeletal proteins during the differentiation of rat mammary tumor cells in vitro

Cited by 39 publications

References 18 publications

Microtubule‐disrupting drugs increase the frequency of conversion of a rat mammary epithelial stem cell line to elongated, myoepithelial‐like cells in culture

Microtubule‐disrupting drugs increase the frequency of conversion of a rat mammary epithelial stem cell line to elongated, myoepithelial‐like cells in culture

Establishment and Characterization of a Buffalo (Bubalus bubalis) Mammary Epithelial Cell Line

Stem cells and the development of mammary cancers in experimental rats and in humans

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